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    • 8. 发明授权
    • Hybridization method and probe for detecting nucleic acid sequences
    • 用于检测核酸序列的杂交方法和探针
    • US4908307A
    • 1990-03-13
    • US944678
    • 1986-12-19
    • Karin D. RodlandPeter J. Russell
    • Karin D. RodlandPeter J. Russell
    • C12Q1/68
    • C12Q1/6832C12Q1/6816
    • Improved hybridization probe compositions and method for detecting nucleic acids of interest are provided. A hybridization probe constructed to contain thionucleotides and a detectable label is hybridized to a nucleotide sequence of interest in the absence of nonpolar solvents, reducing agents and volume exclusion agents. The thionucleotides serve to increase the mass of bound probe molecules, thereby amplifying the signal available to detect the presence of the nucleic acid sequence of interest. Certain cations interact specifically with the thiol moiety of phosphorothioate groups of nucelic acid molecules in a manner that can be exploited to amplify probe binding or convey a detectable label. Thus, the mass of bound probe molecules can be further increased by the presence of specified multivalent cations from the transition metal group, such as Cu.sup.2+ or Fe.sup.3+, during the annealing step.
    • 提供了改进的杂交探针组合物和用于检测感兴趣的核酸的方法。 在不存在非极性溶剂,还原剂和体积排阻剂的情况下,构建成含有硫核苷酸和可检测标记的杂交探针与目的核苷酸序列杂交。 硫代核苷酸用于增加结合的探针分子的质量,从而扩增可用于检测感兴趣的核酸序列的存在的信号。 某些阳离子以可以用于扩增探针结合或传递可检测标记物的方式与硫酸分子的硫醇部分特异性地相互作用。 因此,在退火步骤期间,通过存在来自过渡金属基团(例如Cu 2+或Fe 3+)的特定多价阳离子,结合的探针分子的质量可以进一步增加。